Protocol to measure glucose utilization in mouse tissues using radiolabeled 2-deoxy-D-glucose

• Published in: STAR protocols Vol. 5; no. 4; p. 103478
• Main Authors: Wu, Cyun-Ming, Li, Hao-Yun, Li, Wen-Yi, Hsu, Chih-Neng, Yang, Wei-Shun, Chuang, Gwo-Tsang, Che, Tony Pan-Hou, Lee, Tung-Yuan, Lee, Hsiao-Lin, Hee, Siow-Wey, Lee, Jiin-Horng, Liao, Daniel, Liao, Karen Chia-Wen, Liu, Ya-Wen, Hsuan, Chin-Feng, Chang, Yi-Cheng, Chu, Shao-Lun
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 20.12.2024; Elsevier
• Subjects: Animals; Deoxyglucose - analogs & derivatives; Deoxyglucose - metabolism; Glucose - metabolism; Glucose-6-Phosphate - analogs & derivatives; Health Sciences; Metabolism; Metabolomics; Mice; Protocol; Metabolism; Metabolomics; Health Sciences
• ISSN: 2666-1667; 2666-1667
• DOI: 10.1016/j.xpro.2024.103478

2-deoxy-D-glucose (2DG) is a glucose analog converted to 2-deoxy-D-glucose-6-phosphate (2DG-6P) by hexokinase in glycolysis. While 2DG commonly measures glucose uptake, 2DG-6P detects glucose utilization. Here, we present a protocol to measure glucose utilization in various tissues after entering a mouse’s body using radiolabeled 2DG. We describe steps for preparing mice and chemicals, extracting blood, adding chemicals, and dissolving tissue. We then detail procedures for calculating glucose utilization using the trapezoid rule. [Display omitted] •Use radiolabeled 2DG converted to 2DG-6P to measure glucose utilization in tissues•Steps for performing preparation, extracting blood, adding chemicals, and dissolving tissue•Steps to convert radioactive numbers to glucose utilization using trapezoidal technique Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. 2-deoxy-D-glucose (2DG) is a glucose analog converted to 2-deoxy-D-glucose-6-phosphate (2DG-6P) by hexokinase in glycolysis. While 2DG commonly measures glucose uptake, 2DG-6P detects glucose utilization. Here, we present a protocol to measure glucose utilization in various tissues after entering a mouse’s body using radiolabeled 2DG. We describe steps for preparing mice and chemicals, extracting blood, adding chemicals, and dissolving tissue. We then detail procedures for calculating glucose utilization using the trapezoid rule.